1. Prepare assay buffer in a suitable container and pre-chill on ice before use: The final concentrations are 35 mM Tris, pH 8.0, 1uM suc-AAFP-pNA). 2. Add recombinant PPIL1 protein with various concentrations (1ug, 2ug, 5ug) in assay buffer. 3. Mix by inversion and equilibrate to 1 DEG C and monitor the A410nm until the value is constant using a spectrophotometer. 4. Add pre-chilled chymotrypsin to 12.5 uM and mix immediately. 5. Record the increase in A410 nm for 20 minutes

Usage & Assay Information

Specific activity is > 155 nM/min/ug, and is defined as the amount of enzyme that cleaves 1uM of suc-AAFP-pNA per minute at 1 DEG C in Tris-Hcl pH 8.0 using chymotrypsin.

Storage & Safety

Storage

Store at 4 deg C for short term storage (1/2 weeks). Aliquot and store at -20 deg C or - 70 deg C for long term storage. Avoid repeated freeze/thaw cycles.

General Information

Biological Significance

Peptidylprolyl isomerase (cyclophilin)-like 1, also known as PPIL1, is a member of peptidyl-propyl cis-trans isomerase (PPIase) family, which catalyzes the cis-trans isomerization of proline imidic peptide bonds in oligopeptides and accelerates the folding of proteins. Human PPIL1 might play an important role in proliferation of cancer cells through modulation of phosphorylation of stathmin. So it is expected as a novel molecular target for colon-cancer therapy. Recombinant PPIL1, fused to His-tag at C-terminus, was expressed in E. coli and purified by conventional chromatography techniques.